期刊
AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE
卷 94, 期 6, 页码 1318-1323出版社
AMER SOC TROP MED & HYGIENE
DOI: 10.4269/ajtmh.15-0829
关键词
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资金
- Japan Society for the Promotion of Science (JSPS) [26-04103]
- JSPS [15H05273, 21406009, 24406011, 21256003, 24256002]
- JSPS-Asia/Africa Scientific Platform Fund
- Special Coordination Fund for Promoting Science and Technology from the Ministry of Education, Japan
- Grants-in-Aid for Scientific Research [15H05273, 14F04103, 21406009, 21256003, 24406011] Funding Source: KAKEN
For differential detection of Taenia solium, Taenia saginata, and Taenia asiatica, loop-mediated isothermal amplification (LAMP) assay targeting the cytochrome c oxidase subunit 1 gene has been recently developed and shown to be sensitive, specific, and effective. However, to achieve differential identification, one specimen requires three reaction mixtures containing a primer set of each Taenia species separately, which is complex and time consuming and increases the risk of cross-contamination. In this study, we developed a simple differential identification of human Taenia species using multiplex LAMP (mLAMP) in combination with dot enzyme-linked immunosorbent assay (dot-ELISA). Forward inner primers of T solium, T saginata, and T asiatica labeled with fluorescein isothiocyanate (FITC), digoxigenin (DIG), and tetramethylrhodamine (TAMRA), respectively, and biotin-labeled backward inner primers were used in mLAMP. The mLAMP assay succeeded in specific amplification of each respective target gene in a single tube. Furthermore, the mLAMP product from each species was easily distinguished by dot-ELISA with an antibody specific for FITC, DIG, or TAMRA. The mLAMP assay in combination with dot-ELISA will make identification of human Taenia species simpler, easier, and more practical.
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